EFFECT OF DENERVATION ON THE FUNCTION OF THE AIRWAYS

去神经支配对气道功能的影响

• 批准号: 6184175
• 负责人: Julio PEREZ FONTAN
• 金额: $ 24.3万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6184175
• 关键词: DNA footprinting; SCID mouse; denervation; ganglion cell; gene expression; genetic promoter element; growth factor receptors; human tissue; in situ hybridization; laboratory rat; lung transplantation; messenger RNA; neurons; neurotrophic factors; phenotype; platelet derived growth factor; procollagen; receptor expression; respiratory function; substance K; sympathetic ganglion; tachykinin; transforming growth factors; vagotomy; vagus nerve

This proposal will study the effects of lung denervation on the structure and function of the airways by testing two general hypotheses: a.increased airway strain during breathing initiates a fibroproliferative response in denervated airways and b. neurotrophins produced by denervated neuronal targets modify neuropeptide gene expression in airway parasympathetic ganglia. The experimental approach will pursue two specific aim. The first aim will be to define the effects of denervation on the transciptional regulation of type I procollagen in rats. Accordingly, procollagen expression will be assessed by i situ hybridization after unilateral lung vagotomy or syngeneic lung transplantation. Differences in procollagen mRNA levels between denervated and control lungs will be: 1. examined after experimental manipulations of airway strain and 2. related to the expression of two growth factors which contain strain- responsive elements in their gene promoter and may up-regulate collagen transcription: platelet-derived growth factor (PDGF) and transforming growth factor beta (TGF beta). The second aim will be to elucidate the mechanisms that determine the peptidergic phenotype of denervated human and rat airway parasympathetic ganglia. The expressions of protachykinin gene products and their receptors will be studied by in situ hybridization, solution hybridization-nuclease protection assays, and immunohistochemistry in denervated rat lungs and xenografts prepared by implanting rat airways and human bronchi into subcutaneous tissue of immonodeficient mice. The resultant temporal profile of pre-protachykinin and neurokinin receptor expressions will be: 1. related pharmacologically to neurokinin contribution to bronchomotor tone, 2 evaluated after extended neurokinin receptor inhibition, and 3. contrasted to the expression of neurotrophins by denervated airway tissues and neurotrophin receptors by ganglion neurons. The information gained from this research will not only advance the current understanding of the biological function of airway nerves, but will also provide valuable insight into the pathogenesis of lung transplantation-induced bronchiolitis obliterans and other potential airway injuries cause by pharmacological or mechanical disruption of airway smooth muscle tone.

这项建议将研究肺去神经对结构的影响， 和功能的气道通过测试两个一般假设： 呼吸期间的气道紧张引发纤维增生反应， 去神经气道和B.失神经支配神经元产生的神经营养因子 靶点修饰气道副交感神经中神经肽基因表达 神经节 实验方法将追求两个具体目标。 的 第一个目标是确定去神经支配对 大鼠I型前胶原的转录调控。 因此，委员会认为， 前胶原表达将通过原位杂交评估， 单侧肺迷走神经切断术或同基因肺移植。 差异 去神经支配肺和对照肺之间的前胶原mRNA水平将是： 1.检查后，实验操作的气道应变和2。 与两种生长因子的表达有关， 在其基因启动子中的响应元件，并可能上调胶原蛋白 转录：血小板衍生生长因子（PDGF）和转化 生长因子β（TGF β）。 第二个目标是阐明 决定失神经支配的人的肽能表型和 大鼠气道副交感神经节。 前速激肽基因的表达 产物及其受体将通过原位杂交进行研究， 溶液杂交-核酸酶保护测定和免疫组织化学 在去神经大鼠肺和通过植入大鼠气道制备的异种移植物中， 和人支气管进入免疫缺陷小鼠的皮下组织。 的 前速激肽原和神经激肽受体的合成时间分布 表达式将是：1.与神经激肽相关的神经递质 对支气管张力的贡献，2在延长神经激肽后评价 受体抑制，和3.与神经营养因子的表达相比， 通过去神经支配的气道组织和神经营养因子受体通过神经节 神经元从这项研究中获得的信息不仅会促进 目前对气道神经生物学功能的理解，但 也将提供有价值的洞察肺的发病机制， 移植诱导的闭塞性细支气管炎和其他潜在气道 由于药物或机械性破坏气道平滑而造成的损伤 肌肉张力